HEK293S GnTI cell line细胞株 BioVector NTCC质粒载体菌种细胞基因保藏中心

HEK293S GnTI cell line细胞株

Organism  Homo sapiens, human
Tissue  embryonic kidney
Cell Type  transformed with adenovirus 5 DNA
Product Format  frozen
Morphology  epithelial-like
Culture Properties  adherent
Biosafety Level  2   [Cells contain adenovirus viral DNA sequences]

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Age  fetus
Applications  
Widely used to overexpress various mammalian membrane proteins.
Genes Expressed  
N-acetylglucosaminyltransferase I (GnTI), not expressed
Ref
Cellular Products  
N-acetylglucosaminyltransferase I (GnTI), not expressed
Ref
Comments  

The HEK293S GnTIˉ cell line was established by methanesulfonate mutagenesis followed by Ricin selection. HEK293S GnTIˉ cells do not have N-acetyl-glucosaminyltransferase I (GnTI) activity and therefore lack complex N-glycans.Ref  This cell line has been successfully used to overexpress a wide variety of mammalian membrane proteins. There were two versions of HEK293S GnTIˉ cells in the original publication. One was HEK293S GnTIˉ (ATCC CRL-3022), and the other version was HEK293S GnTIˉ variant constructed with tetracycline-inducible system. CRL-3022 does not contain tetracycline-inducible system or pcDNA6-TR vector.  
Complete Growth Medium  The base medium for this cell line is ATCC-formulated DMEM:F12 Medium Catalog No. 30-2006. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Subculturing  
These cells adhere poorly to standard tissue culture flasks. Attachment can be enhanced by using Corning CellBIND® flasks.

Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

   Remove and discard culture medium.
   Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
   Add 1.0 to 2.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
   Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
   Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
   Add appropriate aliquots of the cell suspension to new culture vessels.
   Incubate cultures at 37°C.


Subcultivation ratio: A subcultivation ratio of 1:5 to 1:20 is recommended.
Medium renewal: every 2 to 3 days
Cryopreservation  
Freeze medium: complete growth medium, 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions  
Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5%

Supplier来源:BioVector NTCC Inc.
TEL电话:+86-010-53513060
Website网址: http://www.biovector.net